Collagen III Recombinant Rabbit Monoclonal Antibody [JE60-79]
cat.: HA720050
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: JE60-79
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 139 kDa
Isotype: IgG
Immunogen: Recombinant protein within human Collagen III 101-400/1,466.
Positive control: HeLa cell lysate, A431 cell lysate, SiHa cell lysate, HaCaT cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, C6 cell lysate, PC-12 cell lysate, Mouse heart tissue lysate, Mouse skin tissue lysate, Mouse skeletal muscle tissue lysate, Rat skeletal muscle tissue lysate, Rat skin tissue lysate, SiHa, HeLa.
Subcellular location: Extracellular matrix.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:500-1:2,000
1:50-1:100
1:500-1:1,000
Uniprot #: SwissProt: P02461 Human | P08121 Mouse | P13941 Rat
Alternative names: Alpha 1 type III collagen Alpha1 (III) collagen CO3A1_HUMAN COL 3A1 COL3A1 Collagen alpha 1(III) chain Collagen alpha-1(III) chain Collagen III alpha 1 chain precursor Collagen III alpha 1 polypeptide Collagen type III alpha 1 (Ehlers Danlos syndrome type IV autosomal dominant) Collagen type III alpha 1 Collagen type III alpha 1 chain Collagen type III alpha Collagen, fetal EDS4A Ehlers Danlos syndrome type IV, autosomal dominant Fetal collagen Type III collagen
Images
HA720050_1.jpg Fig1: Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (HA720050) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A431 cell lysate (20 µg/Lane)
Lane 3: SiHa cell lysate (20 µg/Lane)
Lane 4: HaCaT cell lysate (20 µg/Lane)
Lane 5: NIH/3T3 cell lysate (20 µg/Lane)
Lane 6: RAW264.7 cell lysate (20 µg/Lane)
Lane 7: C6 cell lysate (20 µg/Lane)
Lane 8: PC-12 cell lysate (20 µg/Lane)
Lane 9: Mouse heart tissue lysate (20 µg/Lane)
Lane 10: Mouse skin tissue lysate (20 µg/Lane)

Predicted band size: 139 kDa
Observed band size: 150 kDa

Exposure time: 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720050) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA720050_2.jpg Fig2: Western blot analysis of Collagen III on different lysates with Rabbit anti-Collagen III antibody (HA720050) at 1/1,000 dilution.

Lane 1: Mouse heart tissue lysate (20 µg/Lane)
Lane 2: Mouse skeletal muscle tissue lysate (20 µg/Lane)
Lane 3: Rat skeletal muscle tissue lysate (20 µg/Lane)
Lane 4: Rat skin tissue lysate (20 µg/Lane)

Predicted band size: 139 kDa
Observed band size: 150 kDa

Exposure time: 2 minutes; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA720050) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA720050_3.jpg Fig3: Immunocytochemistry analysis of SiHa cells labeling Collagen III with Rabbit anti-Collagen III antibody (HA720050) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (HA720050) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA720050_4.jpg Fig4: Immunocytochemistry analysis of PC-12 cells labeling Collagen III with Rabbit anti-Collagen III antibody (HA720050) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Collagen III antibody (HA720050) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA720050_5.jpg Fig5: Flow cytometric analysis of HeLa cells labeling Collagen III.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA720050, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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